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Genomic Complexity

Genomic Complexity. How many genes in the chromosome?. DNA or RNA? Double Stranded or Single Stranded? Shape? Number of copies? Size in base pairs? Assume 400 amino acids per protein, then how many bases per gene for a typical protein? How many genes possible? Do genes overlap?

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Genomic Complexity

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  1. Genomic Complexity

  2. How many genes in the chromosome? • DNA or RNA? • Double Stranded or Single Stranded? • Shape? • Number of copies? • Size in base pairs? • Assume 400 amino acids per protein, then how many bases per gene for a typical protein? • How many genes possible? • Do genes overlap? • Are both strands transcribed?

  3. Consider the Ribosome

  4. And its operon

  5. Studying Macromolecules –Extracting DNA Cell Pellet Random fragments ~20-40 kbp

  6. Extraction of Chromosomal DNA • DNeasy Tissue Kit • Chromosomal DNA, ~ 20-40 kbp size fragments • Kim Foster 23 kbp Chromosomal DNA RNA Chromosomal DNA RNA

  7. Hybridization

  8. Quantitative Hybridization

  9. Liquid Scintillation Counters Photomultiplier Tubes Electronic Noise Spike 3H 14C 32P log Scale

  10. Experiment 1 – Total 14C-Uracil 10 µg RNA 5 µg RNA sticks 10 µg DNA 5 µg RNA washes off

  11. Experiment 2 – rRNA14C-Uracil 10 µg rRNA 35 ngrRNA sticks 10 µg DNA >99% rRNA washes off

  12. Experiment 3 – tRNA14C-Uracil 10 µg tRNA 5 ngtRNA sticks 10 µg DNA >99% tRNA does not stick

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